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Flow Cytometry Has a Significant Impact on the Cellular Metabolome

Binek, Aleksandra, Rojo, David, Godzien, Joanna, Rupérez, Francisco Javier, Nuñez, Vanessa, Jorge, Inmaculada, Ricote, Mercedes, Vázquez, Jesús, Barbas, Coral·2018·Journal of Proteome Research
DOI 10.1021/acs.jproteome.8b00472
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Abstract

The characterization of specialized cell subpopulations in a heterogeneous tissue is essential for understanding organ function in health and disease. A popular method of cell isolation is fluorescence-activated cell sorting (FACS) based on probes that bind surface or intracellular markers. In this study, we analyze the impact of FACS on the cell metabolome of mouse peritoneal macrophages. Compared with directly pelleted macrophages, FACS-treated cells had an altered content of metabolites related to the plasma membrane, activating a mechanosensory signaling cascade causing inflammation-like stress. The procedure also triggered alterations related to energy consumption and cell damage. The observed changes mostly derive from the physical impact on cells during their passage through the instrument. These findings provide evidence of FACS-induced biochemical changes, which should be taken into account in the design of robust metabolic assays of cells separated by flow cytometry.

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Metadata

Authors
Binek, A., Rojo, D., Godzien, J., Rupérez, F. J., Nuñez, V., Jorge, I., Ricote, M., Vázquez, J., & Barbas, C.
Affiliations
  • Fundación Centro Nacional de Investigaciones Cardiovasculares Carlos III, Madrid 28029, Spain1
  • Centro de Metabolómica y Bioanálisis (CEMBIO), Facultad de Farmacia, Universidad CEU San Pablo, Campus Montepríncipe, Madrid 28668, Spain
ORCID
Venue
Journal of Proteome Research
Publisher
American Chemical Society (ACS)
Published
2018-11-02
ISSN
  • 1535-3893
  • 1535-3907
DOI
10.1021/acs.jproteome.8b00472
Cited by
36
References
2
Language
en
Funders
  • FP7 People: Marie-Curie Actions (608027)
  • Ministerio de Ciencia e Innovaci?n (SEV-2015-0505)
  • Ministerio de Econom?a y Competitividad (SAF2015-71878-REDT, CTQ2014-55279-R, SAF2015-64287R)
  • Instituto de Salud Carlos III (BIO2015-67580-P)
  • Fundaci? la Marat? de TV3 (201605-30-31-32)

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